LOCUS       sequence-297809-        6573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6573)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6573)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046704
FEATURES             Location/Qualifiers
     source          1..6573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(2328..2346)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2697..3500
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      3583..4565
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4643..4672
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4694..5281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5293..5320)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5344..5535)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5536..6327,6328..6396))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6159..6178
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6397..6499)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6500..6531)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 gcagtcggcg cttgtgtatt atgcaactcc cagacgtcgt tgagatgtgg tgcttgtatt
      541 cgtagacctt tcctttgctg taaatgttgc tatgaccacg ttatctcaac ttctcataag
      601 ctagttctga gcgtcaatcc atacgtgtgt aacgcccccg ggtgtgatgt tacagacgtt
      661 acccaattgt acttgggagg tatgtcttac tattgtaagt cccacaagcc acctatatct
      721 tttccattat gcgctaacgg tcaagtcttc ggtttgtaca aaaatacctg tgttggtagt
      781 gataatgtta ccgactttaa cgccattgct acttgtgatt ggactaacgc gggtgattat
      841 atcttggcta acacttgtac cgagaggctt aagttgttcg ccgctgaaac cttaaaggct
      901 acggaagaaa cattcaaatt gagttacggt atcgctactg tcagagaagt tctatctgat
      961 agagaattgc atctctcctg ggaggtgggt aagccaagac caccgttgaa ccggaattac
     1021 gtctttactg gatacagggt tactaagaac tctaaggttc aaattgggga atataccttt
     1081 gaaaagggcg actacggtga cgccgtagtc tatagaggta ccaccactta caaattgaac
     1141 gtcggtgact atttcgtttt aaccagccac acagtgatgc cactgtcagc gccaacatta
     1201 gttccgcaag agcattacgt tagaattact ggtttgtatc caactctaaa tatctctgat
     1261 gaattttcct ccaatgtggc taactaccaa aaagtaggta tgcaaaagta ctctaccttg
     1321 caaggtccac ctggtactgg gaagtcgcac ttcgctatag gtttggcact gtactaccca
     1381 tctgctagaa ttgtttacac ggcatgttct catgctgctg tagacgcctt gtgtgaaaag
     1441 gccttgaaat atttacccat tgataaatgt tcaagaatta tcccagcacg agctcgtgtt
     1501 gaatgtttcg acaagtttaa ggtcaacagc acactagaac aatacgtctt ctgcactgtt
     1561 aatgctttgc cagaaacgac tgctgatatc gtcgtctttg atgaaatctc gatggccact
     1621 aactatgact tatccgtagt taatgctaga cttcgtgcaa agcactacgt ctatattggt
     1681 gatccagctc agctgcctgc accaagaact ttgttaacca agggtacctt ggaaccagaa
     1741 tacttcaact ctgtgtgtcg cctgatgaag acaatcggcc ctgatatgtt cctaggtaca
     1801 tgtagacgtt gtccagctga aattgtggac acggtcagtg ccttggttta cgataacaaa
     1861 ttaaaggcgc ataaggacaa aagcgctcaa tgtttcaaga tgttttacaa gggtgtcatt
     1921 actcacgatg ttagttcggc catcaataga ccccaaatag gtgttgttag agaatttttg
     1981 acaaggaacc cagcatggag aaaggctgtt ttcatatccc catacaattc tcaaaacgcg
     2041 gtagcctcaa aaatcttggg tcttcccact cagaccgttg attcttccca aggaagtgaa
     2101 tatgactacg tgattttcac tcaaaccacc gagactgccc actcttgcaa cgtcaatcgt
     2161 ttcaacgttg ctattacgag agccaaggtt ggcatcttgt gtatcatgtc tgatagagac
     2221 ttgtacgata aattacagtt tacatcctta gaaataccaa gacgaaacgt cgctactttg
     2281 caataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat
     2341 tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     2401 taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta tttatatttc
     2461 aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac tgcagtacac
     2521 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
     2581 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
     2641 cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt
     2701 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
     2761 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
     2821 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
     2881 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
     2941 ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt aatacagtca
     3001 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
     3061 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
     3121 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
     3181 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
     3241 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     3301 ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga accgtggatg
     3361 atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     3421 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     3481 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     3541 aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc atctgtgctt
     3601 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     3661 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     3721 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     3781 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa
     3841 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     3901 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     3961 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     4021 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     4081 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     4141 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     4201 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     4261 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     4321 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     4381 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     4441 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt
     4501 ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt
     4561 tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta
     4621 actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa
     4681 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4741 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4801 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4861 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4921 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4981 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5041 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5101 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5161 gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5221 ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5281 aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc
     5341 tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     5401 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     5461 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     5521 ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5581 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tgcgggaggg
     5641 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5701 tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc ctgcaacttt
     5761 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5821 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5881 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5941 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa gtaagttggc
     6001 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6061 cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6121 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6181 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct caaggatctt
     6241 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6301 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6361 gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag ctcctgaaaa
     6421 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     6481 acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat
     6541 ggaggtcagg tatgatttaa atggtcagtt aga
//
